PDF(Pubmed)
Abstract：
BACKGROUND: Staphylococcus aureus is a notorious multidrug resistant pathogen prevalent in healthcare facilities worldwide. Unveiling the mechanisms underlying biofilm formation, quorum sensing and antibiotic resistance can help in developing more effective therapy for S. aureus infection. There is a scarcity of literature addressing the genetic profiles and correlations of biofilm-associated genes, quorum sensing, and antibiotic resistance among S. aureus isolates from Malaysia.
METHODS: Biofilm and slime production of 68 methicillin-susceptible S. aureus (MSSA) and 54 methicillin-resistant (MRSA) isolates were determined using a a plate-based crystal violet assay and Congo Red agar method, respectively. The minimum inhibitory concentration values against 14 antibiotics were determined using VITEK® AST-GP67 cards and interpreted according to CLSI-M100 guidelines. Genetic profiling of 11 S. aureus biofilm-associated genes and agr/sar quorum sensing genes was performed using single or multiplex polymerase chain reaction (PCR) assays.
RESULTS: In this study, 75.9% (n = 41) of MRSA and 83.8% (n = 57) of MSSA isolates showed strong biofilm-forming capabilities. Intermediate slime production was detected in approximately 70% of the isolates. Compared to MSSA, significantly higher resistance of clindamycin, erythromycin, and fluoroquinolones was noted among the MRSA isolates. The presence of intracellular adhesion A (icaA) gene was detected in all S. aureus isolates. All MSSA isolates harbored the laminin-binding protein (eno) gene, while all MRSA isolates harbored intracellular adhesion D (icaD), clumping factors A and B (clfA and clfB) genes. The presence of agrI and elastin-binding protein (ebpS) genes was significantly associated with biofilm production in MSSA and MRSA isolates, respectively. In addition, agrI gene was also significantly correlated with oxacillin, cefoxitin, and fluoroquinolone resistance.
CONCLUSIONS: The high prevalence of biofilm and slime production among MSSA and MRSA isolates correlates well with the detection of a high prevalence of biofilm-associated genes and agr quorum sensing system. A significant association of agrI gene was found with cefoxitin, oxacillin, and fluoroquinolone resistance. A more focused approach targeting biofilm-associated and quorum sensing genes is important in developing new surveillance and treatment strategies against S. aureus biofilm infection.
METHODS: Biofilm and slime production of 68 methicillin-susceptible S. aureus (MSSA) and 54 methicillin-resistant (MRSA) isolates were determined using a a plate-based crystal violet assay and Congo Red agar method, respectively. The minimum inhibitory concentration values against 14 antibiotics were determined using VITEK® AST-GP67 cards and interpreted according to CLSI-M100 guidelines. Genetic profiling of 11 S. aureus biofilm-associated genes and agr/sar quorum sensing genes was performed using single or multiplex polymerase chain reaction (PCR) assays.
RESULTS: In this study, 75.9% (n = 41) of MRSA and 83.8% (n = 57) of MSSA isolates showed strong biofilm-forming capabilities. Intermediate slime production was detected in approximately 70% of the isolates. Compared to MSSA, significantly higher resistance of clindamycin, erythromycin, and fluoroquinolones was noted among the MRSA isolates. The presence of intracellular adhesion A (icaA) gene was detected in all S. aureus isolates. All MSSA isolates harbored the laminin-binding protein (eno) gene, while all MRSA isolates harbored intracellular adhesion D (icaD), clumping factors A and B (clfA and clfB) genes. The presence of agrI and elastin-binding protein (ebpS) genes was significantly associated with biofilm production in MSSA and MRSA isolates, respectively. In addition, agrI gene was also significantly correlated with oxacillin, cefoxitin, and fluoroquinolone resistance.
CONCLUSIONS: The high prevalence of biofilm and slime production among MSSA and MRSA isolates correlates well with the detection of a high prevalence of biofilm-associated genes and agr quorum sensing system. A significant association of agrI gene was found with cefoxitin, oxacillin, and fluoroquinolone resistance. A more focused approach targeting biofilm-associated and quorum sensing genes is important in developing new surveillance and treatment strategies against S. aureus biofilm infection.
摘要：
背景：金黄色葡萄球菌是全球医疗机构中普遍存在的臭名昭著的多重耐药病原体。揭开生物膜形成的潜在机制，群体感应和抗生素耐药性可以帮助开发更有效的金黄色葡萄球菌感染治疗方法。缺乏涉及生物膜相关基因的遗传概况和相关性的文献，仲裁感应,以及来自马来西亚的金黄色葡萄球菌分离株的抗生素耐药性。
方法：使用基于平板的结晶紫测定法和刚果红琼脂法测定了68种甲氧西林敏感的金黄色葡萄球菌（MSSA）和54种耐甲氧西林（MRSA）分离株的生物膜和粘液的产生，分别。使用VITEK®AST-GP67卡确定针对14种抗生素的最小抑制浓度值,并根据CLSI-M100指南进行解释。使用单一或多重聚合酶链反应（PCR）测定对11个金黄色葡萄球菌生物膜相关基因和agr/sar群体感应基因进行遗传分析。
结果：在这项研究中，75.9％（n=41）的MRSA和83.8％（n=57）的MSSA分离株显示出强的生物膜形成能力。在大约70％的分离物中检测到中间粘液的产生。与MSSA相比，克林霉素的耐药性明显更高，红霉素,MRSA分离株中注意到氟喹诺酮类药物。在所有金黄色葡萄球菌分离物中检测到细胞内粘附A（icaA）基因的存在。所有MSSA分离物都含有层粘连蛋白结合蛋白（eno）基因，而所有MRSA分离株都具有细胞内粘附性D(icaD)，聚集因子A和B(clfA和clfB)基因。在MSSA和MRSA分离株中，agrI和弹性蛋白结合蛋白（ebpS）基因的存在与生物膜的产生显着相关，分别。此外,agrI基因也与苯唑西林显著相关，头孢西丁,和氟喹诺酮耐药。
结论：MSSA和MRSA分离株中生物膜和粘液产生的高流行率与生物膜相关基因和agr群体感应系统的高流行率密切相关。发现agrI基因与头孢西丁有显著关联，苯唑西林,和氟喹诺酮耐药。靶向生物膜相关和群体感应基因的更集中的方法在开发针对金黄色葡萄球菌生物膜感染的新监测和治疗策略中是重要的。
方法：使用基于平板的结晶紫测定法和刚果红琼脂法测定了68种甲氧西林敏感的金黄色葡萄球菌（MSSA）和54种耐甲氧西林（MRSA）分离株的生物膜和粘液的产生，分别。使用VITEK®AST-GP67卡确定针对14种抗生素的最小抑制浓度值,并根据CLSI-M100指南进行解释。使用单一或多重聚合酶链反应（PCR）测定对11个金黄色葡萄球菌生物膜相关基因和agr/sar群体感应基因进行遗传分析。
结果：在这项研究中，75.9％（n=41）的MRSA和83.8％（n=57）的MSSA分离株显示出强的生物膜形成能力。在大约70％的分离物中检测到中间粘液的产生。与MSSA相比，克林霉素的耐药性明显更高，红霉素,MRSA分离株中注意到氟喹诺酮类药物。在所有金黄色葡萄球菌分离物中检测到细胞内粘附A（icaA）基因的存在。所有MSSA分离物都含有层粘连蛋白结合蛋白（eno）基因，而所有MRSA分离株都具有细胞内粘附性D(icaD)，聚集因子A和B(clfA和clfB)基因。在MSSA和MRSA分离株中，agrI和弹性蛋白结合蛋白（ebpS）基因的存在与生物膜的产生显着相关，分别。此外,agrI基因也与苯唑西林显著相关，头孢西丁,和氟喹诺酮耐药。
结论：MSSA和MRSA分离株中生物膜和粘液产生的高流行率与生物膜相关基因和agr群体感应系统的高流行率密切相关。发现agrI基因与头孢西丁有显著关联，苯唑西林,和氟喹诺酮耐药。靶向生物膜相关和群体感应基因的更集中的方法在开发针对金黄色葡萄球菌生物膜感染的新监测和治疗策略中是重要的。
